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Literature summary for 3.2.1.176 extracted from

  • Voutilainen, S.P.; Boer, H.; Alapuranen, M.; Jaenis, J.; Vehmaanperae, J.; Koivula, A.
    Improving the thermostability and activity of Melanocarpus albomyces cellobiohydrolase Cel7B (2009), Appl. Microbiol. Biotechnol., 83, 261-272 .
    View publication on PubMed
Search for more literature for 3.2.1.176

Cloned(Commentary)

Cloned (Comment) Organism
expression in Saccharomyces cerevisiae Melanocarpus albomyces

Protein Variants

Protein Variants Comment Organism
G4C/M70C introduction of disulfide bridge, improvement of thermostability by 2.5 degrees and increased activity towards microcrystalline cellulose at 75°C Melanocarpus albomyces
G4C/M70C/S290T improvement of thermostability by 4.0 degrees and increased activity towards microcrystalline cellulose at 75°C Melanocarpus albomyces
additional information linking of a cellulose-binding module improves the thermostability by 2.5 degrees and causes a clear increase in the hydrolysis activity towards Avicel at 70°C Melanocarpus albomyces
S290T improvement of thermostability by 1.5 degrees Melanocarpus albomyces

Inhibitors

Inhibitors Comment Organism Structure
cellobiose mixed type inhibition Melanocarpus albomyces

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.23
-
 4-methylumbelliferyl beta-D-lactopyranoside wild-type, pH 6.0, 22°C Melanocarpus albomyces
0.28
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant S290T, pH 6.0, 22°C Melanocarpus albomyces
0.3
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant G4C/M70C, pH 6.0, 22°C Melanocarpus albomyces
0.3
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant G4C/M70C/S290T, pH 6.0, 22°C Melanocarpus albomyces
0.62
-
 2-chloro-4-nitrophenyl beta-D-lactoside wild-type, pH 6.0, 22°C Melanocarpus albomyces
0.75
-
 2-chloro-4-nitrophenyl beta-D-lactoside mutant G4C/M70C/S290T, pH 6.0, 22°C Melanocarpus albomyces
1
-
 2-chloro-4-nitrophenyl beta-D-lactoside mutant G4C/M70C, pH 6.0, 22°C Melanocarpus albomyces
1.16
-
 2-chloro-4-nitrophenyl beta-D-lactoside mutant S290T, pH 6.0, 22°C Melanocarpus albomyces

Organism

Organism UniProt Comment Textmining
Melanocarpus albomyces Q8J0K6
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-chloro-4-nitrophenyl beta-D-lactoside + H2O
-
 Melanocarpus albomyces 2-chloro-4-nitrophenol + D-lactose
-
 ?
4-methylumbelliferyl beta-D-lactopyranoside + H2O
-
 Melanocarpus albomyces 4-methylumbelliferone + lactose
-
 ?

Subunits

Subunits Comment Organism
? x * 47500, calculated, x * 50000, SDS-PAGE, x * 65000, SDS-PAGE of recombinant protein carrying a cellulose-binding module Melanocarpus albomyces

Synonyms

Synonyms Comment Organism
Cel7B
-
 Melanocarpus albomyces

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
68
-
 melting temperature, wild-type Melanocarpus albomyces
70
-
 melting temperature, mutant S290T Melanocarpus albomyces
70.5
-
 melting temperature, mutant carrying a cellulose-binding module Melanocarpus albomyces
71
-
 melting temperature, mutant G4C/M70C Melanocarpus albomyces
72.5
-
 melting temperature, mutant G4C/M70C/S290T Melanocarpus albomyces

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.017
-
 2-chloro-4-nitrophenyl beta-D-lactoside wild-type, pH 6.0, 22°C Melanocarpus albomyces
0.02
-
 2-chloro-4-nitrophenyl beta-D-lactoside mutant S290T, pH 6.0, 22°C Melanocarpus albomyces
0.08
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant G4C/M70C, pH 6.0, 22°C Melanocarpus albomyces
0.11
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant S290T, pH 6.0, 22°C Melanocarpus albomyces
0.12
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant G4C/M70C/S290T, pH 6.0, 22°C Melanocarpus albomyces
0.14
-
 4-methylumbelliferyl beta-D-lactopyranoside wild-type, pH 6.0, 22°C Melanocarpus albomyces
0.165
-
 2-chloro-4-nitrophenyl beta-D-lactoside mutant G4C/M70C/S290T, pH 6.0, 22°C Melanocarpus albomyces
0.182
-
 2-chloro-4-nitrophenyl beta-D-lactoside mutant G4C/M70C, pH 6.0, 22°C Melanocarpus albomyces

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.005
-
 cellobiose mutant G4C/M70C/S290T, pH 6.0, 22°C Melanocarpus albomyces
0.0055
-
 cellobiose mutant G4C/M70C, pH 6.0, 22°C Melanocarpus albomyces
0.006
-
 cellobiose wild-type, pH 6.0, 22°C Melanocarpus albomyces
0.0066
-
 cellobiose mutant S290T, pH 6.0, 22°C Melanocarpus albomyces

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.017
-
 2-chloro-4-nitrophenyl beta-D-lactoside mutant G4C/M70C, pH 6.0, 22°C Melanocarpus albomyces
0.017
-
 2-chloro-4-nitrophenyl beta-D-lactoside mutant S290T, pH 6.0, 22°C Melanocarpus albomyces
0.022
-
 2-chloro-4-nitrophenyl beta-D-lactoside mutant G4C/M70C/S290T, pH 6.0, 22°C Melanocarpus albomyces
0.027
-
 2-chloro-4-nitrophenyl beta-D-lactoside wild-type, pH 6.0, 22°C Melanocarpus albomyces
0.27
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant G4C/M70C, pH 6.0, 22°C Melanocarpus albomyces
0.38
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant S290T, pH 6.0, 22°C Melanocarpus albomyces
0.4
-
 4-methylumbelliferyl beta-D-lactopyranoside mutant G4C/M70C/S290T, pH 6.0, 22°C Melanocarpus albomyces
0.62
-
 4-methylumbelliferyl beta-D-lactopyranoside wild-type, pH 6.0, 22°C Melanocarpus albomyces